Genetic resources of Cannabis sativa L. at the

Genetic resources of Cannabis sativa L. at the Institute of Natural Fibres and
Medicinal Plants in Poznań
Grażyna Mańkowska*, Lidia Grabowska
Institute of Natural Fibres and Medicinal Plants
Wojska Polskiego 71B
60-630 Poznań, Poland
*corresponding author: e-mail: [email protected]
Summary
Hemp (Cannabis sativa L.) species belongs to the Hemp family (Cannabaceae), Cannabis genus.
Hemp is considered as a species easily adapting to the changing environment conditions.
It is characterized by plentiful of forms showing considerable biological and morphological differences and different economical value. Development of such big differentiation
took place under the effect of geographical, climatic and soil conditions as well as human
activity which aimed at developing and stabilizing important qualities of hemp. The collection of hemp genus held at the Institute of Natural Fibres in Poznań contains about 150
accessions. The collection holds both mono- and dioecious forms of hemp from zones
represented by different soil and climatic conditions, local ecotypes and breeding lines
with stabilized genotype that are exceptional in respect of one parameter, for instance
Δ9THC, yield, quality of fiber, composition of fatty acids, etc.
In 2000, the INF genetic resources were transferred to new facilities providing excellent
conditions of storage. The long-term storage of seeds requires low air temperature, low
seed moisture content (7–8%) and proper relative air humidity (below 55%). The air-dried
seeds (7% of moisture content) are packed hermetically in a thermo-sealed aluminum foil
and sent to core collection stored in a chamber at +4.0oC or to reserve bank where the 5-g
samples are stored for a long time.
Key words: genetic resources, monoecious hemp, dioecious hemp, accession, collection, accession
evaluation
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INTRODUCTION
Hemp (Cannabis sativa L.) belongs to the Cannabaceae family, Cannabis genus.
Hemp plants are considered to be readily adapting to changing environmental
conditions. Hemp is characterized by a multitude of diverse forms of biological
and morphological traits and the different economic value [1, 2]. The formation
of such a large difference occurred as a result of geographical, climatic and soil
conditions and human activities, which aimed at establishing and maintaining
important features of commercial hemp.
Currently, the Institute of Natural Fibres and Medicinal Plants holds a collection
consisting of 150 genotypes, including monoecious and dioecious forms of hemp,
from different regions of the world, local ecotypes and breeding lines with a stable
genotype, unique in terms of at least one parameter, such as the THC content,
yield, fiber quality, fatty acids composition etc.
Evaluation work is carried out on collected objects, aimed at renewal and multiplication of seed and allows possession to carry out the work of valorization. For test objects,
in addition to distinctive features, morphological and biological traits and use value
were determined. This data is valuable for breeders in selecting parental components.
MATERIAL AND METODS
Renewal of sowing and the multiplication of genotypes.
Propagation and renewal of sowing material in the collection is conducted at
the Experimental Station of Institute of Natural Fibres and Medicinal Plants in Pętkowo (Wielkopolska province). Due to dioeciousness, allogamy and anemophily
of hemp, the work related to the propagation and renewal of sowing material requires spatial isolation. Small batches of seed are planted in isolated pots located
in the vegetation hall and larger batches of seed in isolated chambers, greenhouses and with the isolation in the field.
In case of reproduction of monoecious forms, it is necessary to remove the
male specimen during flowering. Another problem in case of cannabis is that the
seed very fast loses its germination capacity [3]. Therefore, the seeds of germination capacity higher than 80% are controlled every 3 years, those with germination
capacity lower than 80% – every year.
Determination of the economic value
A series of comparative field experiments was conducted in order to determine
the economic value of the stored genotypes in accordance with the agronomical
recommendations of the hemp fiber. Annually, a bulk of 15–20 objects are tested
in two repetitions of 300 seeds on 1.5 m2 plots.
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During vegetation several observations are carried out e.g. development phases of hemp, lodging and diseases as well as pests and plant number per unit area.
During the flowering inflorescences are sampled in order to determine a hallucinogenic substances.
The following tests were performed in the laboratory:
• measurements of morphological traits: total length, stem length and diameter;
• 1000 seed mass, determined according to ISTA [4];
• determination of fiber content in straw, according to the method of osmotic
degumming developed at INF [5];
• cellulose content, according to PN -92/pn-50092;
• content of hallucinogenic substances – using gas chromatograph in accordance with EU Council Regulation (EC) No 1251/1999, Annex 5, Art. 7b, annex 1.
Białobrzeskie variety serves as a reference cultivar – the first Polish monoecious
fibrous hemp variety, registered in 1968 in the National Register of COBORU, which
contains less than 0.2% of hallucinogenic substances, is a Central European form
adapted to Polish climate and soil conditions. The vegetation period of this variety
is 120–140 days, plant height (depending on cultivation technology) 200–350 cm,
seed yield (depending on the purpose of growing 0.3–1.2 t/ha), straw yield about
10 t/ha fiber yield 2.5–3.0 t/ha [6].
RESULTS AND DISCUSSION
Cellulose fiber content
Processing technology of raw material was carried out at the Laboratory Evaluation of Textile Raw Materials, Institute of Natural Fibers and Medicinal Plants.
The results of the determination of fiber and cellulose content of hemp straw is
shown in table 1. Fiber content in the sites surveyed in 2006–2008 was highly
variable and ranged from 12% for the Chinese variety He Bei to 29.2% for the Hungarian variety Uniko B. The reference variety contains 26.6% of fiber. This value is
exceeded only by 4 varieties: Uniko B, Jus 8, Juso 15, Chamaeleon and Juso 11. The
lowest fiber content was found in a group of accessions with the longest growing
season – He Bei, and varieties with the shortest period of vegetation – Finola.
These results confirm the literature data that indicate that at the flowering phase
of growth of biomass is small but biological processes in stems lead to the rise in
fiber content [7, 8].
The cellulose content in hemp straw demonstrated less variability than the
fiber content. The lowest cellulose content was found in the Bulgarian variety
Silistrenskie (37.7%) and the highest in the Ukrainian variety Jus 8 (46.2%), while
this figure was just 0.3% higher than for the reference.
Genetic resources of Cannabis sativa L. at the Institute of Natural Fibres and Medicinal Plants in Poznań
181
Ta b l e 1 .
The content of fiber in selected varieties of hemp collected in the gene bank in 2006–2008
cultivar
average fibre content [%]
cellulose content [%]
Zenit
23.51
45.38
Juso 11
26.05
43.50
Futura 77
20.49
40.12
Chamaeleon
27.06
44.61
Diana
20.74
39.96
Uniko B
29.16
43.56
Silistrenskie
17.27
37.74
Fedora 17
21.82
45.20
Finola
12.28
40.86
Juso 15
28.36
44.10
K - 195
16.48
40.18
Dnieprowska
19.11
42.77
Fibrimon 24
22.28
45.04
Zolotonoska 26
22.54
44.31
Gigantheus
17.62
40.18
He Bei
12.00
39.32
Jus 8
28.77
46.17
Białobrzeskie
26.60
45.85
1000 seed weight
Special studies were conducted on the quality of selected hemp species. The
indicator of seed size is the measure of thousand seed mass. With regard to the
useful characteristics as well as morphological and biological differences, hemp
can be divided into three groups, namely Northern, Southern and Central European ones. Thousand seed mass for each hemp group is different. Average weight of
1000 seeds for Southern hemp is 18–26 g, intermediate hemp 12–18 g and 8–12 g
for wild hemp [9].
Varieties such as: Silistrenskie, Chamaeleon, Zenit exceeded significantly the
MTS value of the Białobrzeskie variety. Finola variety of all tested cultivars is characterized with the lowest value of the 1000 seed weight (tab. 2).
THC content
The contents of narcotic substances in cannabis – Δ9 THC – (tetrahydrocannabinol) is the result of interaction of both genetics and environmental conditions. Genetic predisposition, environmental conditions, different stages of development and plant fragments can cause significant fluctuations in THC content
(from 0.001 to 0.1%, see tab.2).
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182
THC content has been the highest for Finola variety, which value in our climatic
conditions reaches 0.2% and therefore can be considered as a drug type variety.
Ukrainian varieties reached the lowest value of THC [10].
Ta b l e 2 .
Mass of a thousand seeds THC content (%) in hemp cultivars collected in the Institute of Natural
Fibres and Medicinal Plants in 2006–2008
cultivar
origin
mass of 1000 seeds [g]
THC [%]
Zenit
Romania
18.1
0.015
Juso 11
Ukraine
15.3
0.005
Futura 77
France
12.5
0.045
Chamaeleon
The Netherlands
18.0
Diana
Russia
18.6
0.062
Uniko B
Hungary
17.5
0.009
Silistrenskie
Russia
19.0
0.067
Fedora 17
France
17.3
0.053
Finola
Finland
10.3
0.155
Juso 15
Ukraine
13.3
0.016
K - 195
Poland
18.0
0.044
Dnieprowska
Ukraine
17.1
0.039
Fibrimon 24
France
13.7
0.034
Zolotonoska 26
Ukraine
13.2
0.032
Gigantheus
Italy
15.9
0.047
He Bei
China
17.5
0.008
Jus 8
Ukraine
19.0
0.016
Białobrzeskie
Poland
13.9
0.019
CONCLUSIONS
1. Finola (30.05) and He Bei (1.09) begin flowering at the earliest among the
dioecious varieties of hemp. When it comes to monoecious varieties, the
earliest blooming were Juso11, Juso15 (8.07), and the latest was Futura 77
(Tab. 3).
2. The highest fiber content (29.16%) was reported for the Hungarian variety
Uniko B, while the lowest (12.00%) for the Chinese variety He Bei.
3. The highest content of cellulose has been reported for Jus 8 (46.2%), and the
lowest for Silistrenskie (37.74%).
4. The examined accessions were characterized by the mass of 1000 seed from
19.0 g (Jus 8) to 10.3 g (Finola).
5. Finola variety was characterized by higher content of THC in comparison
with the standard variety.
Genetic resources of Cannabis sativa L. at the Institute of Natural Fibres and Medicinal Plants in Poznań
183
Ta b l e 3 .
Morphological parameters of some hemp cultivars from Institute of Natural Fibres and Medicinal
Plants collection
cultivar
form
beginning of the flowering
full maturity
Zenit
Juso 11
monoecious
13.07
20.09
monoecious
8.07
15.09
Futura 77
monoecious
Chamaeleon
dioecious
Diana
Uniko B
4.08
2.10
28.07
2.10
dioecious
1.07
5.09
dioecious
4.08.
8.10
Silistrenskie
dioecious
10.07
20.09
Fedora 17
monoecious
7.07
15.09
Finola
dioecious
30.05
20.08
Juso 15
monoecious
8.07
15.09
K-195
dioecious
10.07
20.09
Dnieprowska
monoecious
2.07
10.09
Fibrimon 24
monoecious
12.07
20.09
Zolotonosha26
monoecious
20.07
25.09
He Bei
dioecious
1.09
29.09
Can.gigantheus
dioecious
16.08
29.09
Białobrzeskie
monoecious
15.07
20.09
REFERENCES
1. Białousowa J, Bartosik A, Kurhański M, Nagórski A, Tumalewicz B. Konopie, Rośliny włókniste [Fibrous
Plants]. Warszawa 1958:193-323.
2. Bosca I, Karus M. Der Hanfanbau. Heidelberg 1997:195.
3. Ziemnicki Z, Wierzchowiecka K. Wpływ przechowywania nasion konopi w różnych opakowaniach na
zachowanie zdolności kiełkowania. Poznań 1984; 29:99-106.
4. Duczmal K, Tucholska H. Nasiennictwo. Tom 1. Warszawa 2000:316-19.
5. Allam A, Kozłowski K, Konczewicz W. Degumming of fibrous plants based on osmotic pressure
phenomenon. Proceedings of International Conference of FAO European Research Network on Flax and
Allied Fibre Plants for Humanity Welfare, Cairo 2003.
6. Grabowska L, Jaranowska B, Baraniecki P, Tymków J. The Results of Hemp Breeding in Poland. Poznań
1998; 2:103-9.
7. Jaranowska B. Wpływ stopnia dojrzałości konopi jednopiennych uprawianych na glebach mineralnych
na wydajność i jakość włókna. Prace IPWŁ. Poznań 1968; 15:37-54.
8. Chocianowicz W, Cierpucha W. Kurhański M, Szałkowski Z, Waśko J.Poradnik brakarza włókna lnianego
i konopnego. Poznań 1987.
9. Bytnerowicz H. Konopie. Warszawa 1968:19-20.
10. Grabowska L, Mańkowska G, Baraniecki P. Zasoby genowe Cannabis sativa L. w Instytucie Włókien
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G. Mańkowska, L. Grabowska
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Zasoby genowe konopi Cannabis sativa L. w Instytucie Włókien Naturalnych
i Roślin Zielarskich w Poznaniu
Grażyna Mańkowska*, Lidia Grabowska
Instytut Włókien Naturalnych i Roślin Zielarskich
ul. Wojska Polskiego 71B
60-630 Poznań
*autor, do którego należy kierować korespondencję: e-mail: [email protected]
Streszczenie
Konopie (Cannabis sativa L.) są gatunkiem należącym do rodziny konopiowatych Cannabaceae, rodzaju Cannabis. Uważane są za roślinę łatwo adaptującą się do zmiennych warunków środowiskowych. Charakteryzuje się mnóstwem form o zróżnicowanych cechach
biologicznych i morfologicznych oraz o różnej wartości ekonomicznej. Wytworzenie się
tak dużych różnic wystąpiło jako efekt działania warunków geograficznych, klimatycznych
i glebowych oraz działalności człowieka, która miała na celu stworzenie i utrzymanie
ważnych cech konopi. Kolekcja rodzaju konopi w Instytucie Włókien Naturalnych i Roślin
Zielarskich w Poznaniu zawiera około 150 obiektów. Zawiera zarówno jedno- i dwupienne
formy charakterystyczne dla różnych warunków glebowo-klimatycznych, lokalne ekotypy
i linie hodowlane ze stabilnym genotypem, wyjątkowym pod względem jednego parametru, np. zawartości THC, plonowania, jakości włókna, składu kwasów tłuszczowych itp.
W 2000 r. zasoby genowe Instytutu Włókien Naturalnych zostały przeniesione do nowego
miejsca, które zapewnia doskonałe warunki przechowywania nasion. Długoterminowe
przechowywanie wymaga niskiej temperatury, niskiej wilgotności nasion (7–8%) oraz
odpowiedniej względnej wilgotności powietrza (poniżej 55%). Osuszane powietrzem
nasiona (7% wilgotności) są zapakowane hermetycznie w zgrzewaną folię aluminiową
i umieszczane w kolekcji bazowej (przechowywanie w komorze w temperaturze +4,0ºC)
lub banku rezerwowym – próbki 5-gramowe są przechowywane przez dłuższy czas.
Słowa kluczowe: zasoby genowe, konopie jednopienne, dwupienne, obiekty, kolekcja, ocena obiektów